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Mol Cell Biol ; 7(10): 3759-66, 1987 Oct.
Article in English | MEDLINE | ID: mdl-3500398

ABSTRACT

T-cell activation pathways are involved in the regulation of human immunodeficiency virus (HIV) expression. Phorbol 12-myristate 13-acetate (PMA) is a potent inducer of T-cell immune functions and has recently been demonstrated to increase viral replication in cell lines infected with HIV. To define sequences required for viral induction by PMA. T-cell lines were transiently transfected with viral long terminal repeat (LTR) sequences directing chloramphenicol acetyltransferase (CAT) gene expression. PMA added to transfected cell cultures 24 h before harvest reproducibly increased both CAT mRNA and enzyme expression 2- to 2-fold. Sequences necessary for basal and PMA-induced levels of CAT expression were determined by deletion and enhancer reconstitution constructs with fragments and oligonucleotides from the original LTR-CAT expression plasmid. PMA-inducible and basal activity required tandem repeats of a core enhancer element (GGGACTTTCC) located in the LTR between -105 and -82 relative to the RNA start site. The enhancerlike sequence could be inserted at a site distant to the CAT gene open reading frame and functioned in a position- and orientation-independent manner. The data thus define a transcriptionally active regulatory-enhancer element critical to the control of HIV gene expression.


Subject(s)
DNA, Viral/genetics , Enhancer Elements, Genetic , Gene Expression Regulation/drug effects , HIV/genetics , Repetitive Sequences, Nucleic Acid , T-Lymphocytes/microbiology , Tetradecanoylphorbol Acetate/pharmacology , Chromosome Mapping , DNA Mutational Analysis , Transcription, Genetic/drug effects
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